• 제목/요약/키워드: fluorescence microscopy.

검색결과 438건 처리시간 0.036초

Preparation of Core-shell Type Nanoparticles of Poly($\varepsilon$-caprolactone) /Poly(ethylene glycol)/Poly( $\varepsilon$-caprolactone) Triblock Copolymers

  • 류재곤;정영일;김영훈;김인숙;김도훈;김성호
    • Bulletin of the Korean Chemical Society
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    • 제22권5호
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    • pp.467-475
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    • 2001
  • A triblock copolymer based on $poly(\varepsilon-caprolactone)$ (PCL) as the hydrophobic part and poly(ethylene glycol) (PEG) as the hydrophilic portion was synthesized by a ring-opening mechanism of ${\varepsilon}-caprolactone$ with PEG containing a hydroxyl group at bot h ends as an initiator. The synthesized block copolymers of PCL/PEG/PCL (CEC) were confirmed and characterized using various analysis equipment such as 1H NMR, DSC, FT-IR, and WAXD. Core-shell type nanoparticles of CEC triblock copolymers were prepared using a dialysis technique to estimate their potential as a colloidal drug carrier using a hydrophobic drug. From the results of particle size analysis and transmission electron microscopy, the particle size of CEC core-shell type nanoparticles was determined to be about 20-60 nm with a spherical shape. Since CEC block copolymer nanoparticles have a core-shell type micellar structure and small particle size similar to polymeric micelles, CEC block copolymer can self-associate at certain concentrations and the critical association concentration (CAC) was able to be determined by fluorescence probe techniques. The CAC values of the CEC block copolymers were dependent on the PCL block length. In addition, drug loading contents were dependent on the PCL block length: the larger the PCL block length, the higher the drug loading content. Drug release from CEC core-shell type nanoparticles showed an initial burst release for the first 12 hrs followed by pseudo-zero order release kinetics for 2 or 3 days. CEC-2 block copolymer core-shell type nanoparticles were degraded very slowly, suggesting that the drug release kinetics were governed by a diffusion mechanism rather than a degradation mechanism irrelevant to the CEC block copolymer composition.

Generation of a recombinant rabies virus expressing green fluorescent protein for a virus neutralization antibody assay

  • Yang, Dong-Kun;Kim, Ha-Hyun;Park, Yu-Ri;Yoo, Jae Young;Park, Yeseul;Park, Jungwon;Hyun, Bang-Hun
    • Journal of Veterinary Science
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    • 제22권4호
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    • pp.56.1-56.10
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    • 2021
  • Background: Fluorescent antibody virus neutralization (FAVN) test is a standard assay for quantifying rabies virus-neutralizing antibody (VNA) in serum. However, a safer rabies virus (RABV) should be used in the FAVN assay. There is a need for a new method that is economical and time-saving by eliminating the immunostaining step. Objectives: We aimed to improve the traditional FAVN method by rescuing and characterizing a new recombinant RABV expressing green fluorescent protein (GFP). Methods: A new recombinant RABV expressing GFP designated as ERAGS-GFP was rescued using a reverse genetic system. Immuno-fluorescence assay, peroxidase-linked assay, electron microscopy and reverse transcription polymerase chain reaction were performed to confirm the recombinant ERAGS-GFP virus as a RABV expressing the GFP gene. The safety of ERAGS-GFP was evaluated in 4-week-old mice. The rabies VNA titers were measured and compared with conventional FAVN and FAVN-GFP tests using VERO cells. Results: The virus propagated in VERO cells was confirmed as RABV expressing GFP. The ERAGS-GFP showed the highest titer (108.0 TCID50/mL) in VERO cells at 5 days post-inoculation, and GFP expression persisted until passage 30. The body weight of 4-week-old mice inoculated intracranially with ERAGS-GFP continued to increase and the survival rate was 100%. In 62 dog sera, the FAVN-GFP result was significantly correlated with that of conventional FAVN (r = 0.95). Conclusions: We constructed ERAGS-GFP, which could replace the challenge virus standard-11 strain used in FAVN test.

스컬용융법에 의한 CaZrO3:Pr3+ 형광체 합성 및 특성분석 (Synthesis and characterization of CaZrO3:Pr3+ phosphor by skull melting method)

  • 최현민;김영출;석정원
    • 한국결정성장학회지
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    • 제31권5호
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    • pp.228-232
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    • 2021
  • 페로브스카이트 구조를 가지는 Ca1-xZrO3:xPr 형광체를 스컬용융법으로 합성하였다. 합성한 형광체의 결정구조, 형태 및 광학적 특성은 XRD, SEM, 자외선 형광반응 및 광발광을 통해 분석하였다. XRD 측정에서는 CaZrO3:Pr3+의 단결정이 페로브스카이트 구조의 사방정계로 분석되었다. 합성된 형광체는 254 nm의 UV 광에 의해 여기 될 수 있고 방출 스펙트럼 결과는 506, 536 및 548 nm에서 전하전이 3P03H4, 3P13H53P03H5로 인해 CaZrO3:Pr3+는 녹색 발광이 우세하였다.

Optimization of Agrobacterium tumefaciens-Mediated Transformation of Xylaria grammica EL000614, an Endolichenic Fungus Producing Grammicin

  • Jeong, Min-Hye;Kim, Jung A.;Kang, Seogchan;Choi, Eu Ddeum;Kim, Youngmin;Lee, Yerim;Jeon, Mi Jin;Yu, Nan Hee;Park, Ae Ran;Kim, Jin-Cheol;Kim, Soonok;Park, Sook-Young
    • Mycobiology
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    • 제49권5호
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    • pp.491-497
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    • 2021
  • An endolichenic fungus Xylaria grammica EL000614 produces grammicin, a potent nematicidal pyrone derivative that can serve as a new control option for root-knot nematodes. We optimized an Agrobacterium tumefaciens-mediated transformation (ATMT) protocol for X. grammica to support genetic studies. Transformants were successfully generated after co-cultivation of homogenized young mycelia of X. grammica with A. tumefaciens strain AGL-1 carrying a binary vector that contains the bacterial hygromycin B phosphotransferase (hph) gene and the eGFP gene in T-DNA. The resulting transformants were mitotically stable, and PCR analysis showed the integratin of both genes in the genome of transformants. Expression of eGFP was confirmed via fluorescence microscopy. Southern analysis showed that 131 (78.9%) out of 166 transformants contained a single T-DNA insertion. Crucial factors for producing predominantly single T-DNA transformants include 48 h of co-cultivation, pretreatment of A. tumefaciens cells with acetosyringone before co-cultivation, and using freshly prepared mycelia. The established ATMT protocol offers an efficient tool for random insertional mutagenesis and gene transfer in studying the biology and ecology of X. grammica.

증착 온도에 따른 La2MoO6:Dy3+,Eu3+ 형광체 박막의 광학 특성 (Effect of Deposition Temperature on the Optical Properties of La2MoO6:Dy3+,Eu3+ Phosphor Thin Films)

  • 조신호
    • 한국전기전자재료학회논문지
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    • 제32권5호
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    • pp.387-392
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    • 2019
  • $Dy^{3+}$ and $Eu^{3+}$-co-doped $La_2MoO_6$ phosphor thin films were deposited on sapphire substrates by radio-frequency magnetron sputtering at various growth temperatures. The phosphor thin films were characterized using X-ray diffraction (XRD), scanning electron microscopy, ultraviolet-visible spectroscopy, and fluorescence spectrometry. The optical transmittance, absorbance, bandgap, and photoluminescence intensity of the $La_2MoO_6$ phosphor thin films were found to depend on the growth temperature. The XRD patterns demonstrated that all the phosphor thin films, irrespective of growth temperatures, had a tetragonal structure. The phosphor thin film deposited at a growth temperature of $100^{\circ}C$ indicated an average transmittance of 85.3% in the 400~1,100 nm wavelength range and a bandgap energy of 4.31 eV. As the growth temperature increased, the bandgap energy gradually decreased. The emission spectra under ultraviolet excitation at 268 nm exhibited an intense red emission line at 616 nm and a weak emission line at 699 nm due to the $^5D_0{\rightarrow}^7F_2$ and $^5D_0{\rightarrow}^7F_4$ transitions of the $Eu^{3+}$ ions, respectively, and also featured a yellow emission band at 573 nm, resulting from the $^4F_{9/2}{\rightarrow}^6H_{13/2}$ transition of the $Dy^{3+}$ ions. The results suggest that $La_2MoO_6$ phosphor thin films can be used as light-emitting layers for inorganic thin film electroluminescent devices.

An Enzymolysis-Assisted Agrobacterium tumefaciens-Mediated Transformation Method for the Yeast-Like Cells of Tremella fuciformis

  • Wang, Yuanyuan;Xu, Danyun;Sun, Xueyan;Zheng, Lisheng;Chen, Liguo;Ma, Aimin
    • Mycobiology
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    • 제47권1호
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    • pp.59-65
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    • 2019
  • Agrobacterium tumefaciens-mediated transformation (ATMT), as a simple and versatile method, achieves successful transformation in the yeast-like cells (YLCs) of Tremella fuciformis with lower efficiency. Establishment of a more efficient transformation system of YLCs is important for functional genomics research and biotechnological application. In this study, an enzymolysis-assisted ATMT method was developed. The degradation degree of YLCs depends on the concentration and digestion time of Lywallzyme. Lower concentration (${\leq}0.1%$) of Lywallzyme was capable of formation of limited wounds on the surface of YLCs and has less influence on their growth. In addition, there is no significant difference of YLCs growth among groups treated with 0.1% Lywallzyme for different time. The binary vector pGEH under the control of T. fuciformis glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter was utilized to transform the enzymolytic wounded YLCs with different concentrations and digestion time. The results of PCR, Southern blot, quantitative real-time PCR (qRT-PCR) and fluorescence microscopy revealed that the T-DNA was integrated into the YLCs genome, suggesting an efficient enzymolysis-assisted ATMT method of YLCs was established. The highest transformation frequency reached 1200 transformants per $10^6$ YLCs by 0.05% (w/v) Lywallzyme digestion for 15 min, and the transformants were genetically stable. Compared with the mechanical wounding methods, enzymolytic wounding is thought to be a tender, safer and more effective method.

Identification of duck liver-expressed antimicrobial peptide 2 and characterization of its bactericidal activity

  • Hong, Yeojin;Truong, Anh Duc;Lee, Janggeun;Lee, Kyungbaek;Kim, Geun-Bae;Heo, Kang-Nyeong;Lillehoj, Hyun S.;Hong, Yeong Ho
    • Asian-Australasian Journal of Animal Sciences
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    • 제32권7호
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    • pp.1052-1061
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    • 2019
  • Objective: This study was conducted to identify duck liver-expressed antimicrobial peptide 2 (LEAP-2) and demonstrate its antimicrobial activity against various pathogens. Methods: Tissue samples were collected from 6 to 8-week-old Pekin ducks (Anas platyrhynchos domesticus), total RNA was extracted, and cDNA was synthesized. To confirm the duck LEAP-2 transcript expression levels, quantitative real-time polymerase chain reaction was conducted. Two kinds of peptides (a linear peptide and a disulfide-type peptide) were synthesized to compare the antimicrobial activity. Then, antimicrobial activity assay and fluorescence microscopic analysis were conducted to demonstrate duck LEAP-2 bactericidal activity. Results: The duck LEAP-2 peptide sequence showed high identity with those of other avian species (>85%), as well as more than 55% of identity with mammalian sequences. LEAP-2 mRNA was highly expressed in the liver with duodenum next, and then followed by lung, spleen, bursa and jejunum and was the lowest in the muscle. Both of LEAP-2 peptides efficiently killed bacteria, although the disulfide-type LEAP-2 showed more powerful bactericidal activity. Also, gram-positive bacteria was more susceptible to duck LEAP-2 than gram-negative bacteria. Using microscopy, we confirmed that LEAP-2 peptides could kill bacteria by disrupting the bacterial cell envelope. Conclusion: Duck LEAP-2 showed its antimicrobial activity against both gram-positive and gram-negative bacteria. Disulfide bonds were important for the powerful killing effect by disrupting the bacterial cell envelope. Therefore, duck LEAP-2 can be used for effective antibiotics alternatives.

Performance Evaluation of Biozentech Malaria Scanner in Plasmodium knowlesi and P. falciparum as a New Diagnostic Tool

  • Firdaus, Egy Rahman;Park, Ji-Hoon;Muh, Fauzi;Lee, Seong-Kyun;Han, Jin-Hee;Lim, Chae-Seung;Na, Sung-Hun;Park, Won Sun;Park, Jeong-Hyun;Han, Eun-Taek
    • Parasites, Hosts and Diseases
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    • 제59권2호
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    • pp.113-119
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    • 2021
  • The computer vision diagnostic approach currently generates several malaria diagnostic tools. It enhances the accessible and straightforward diagnostics that necessary for clinics and health centers in malaria-endemic areas. A new computer malaria diagnostics tool called the malaria scanner was used to investigate living malaria parasites with easy sample preparation, fast and user-friendly. The cultured Plasmodium parasites were used to confirm the sensitivity of this technique then compared to fluorescence-activated cell sorting (FACS) analysis and light microscopic examination. The measured percentage of parasitemia by the malaria scanner revealed higher precision than microscopy and was similar to FACS. The coefficients of variation of this technique were 1.2-6.7% for Plasmodium knowlesi and 0.3-4.8% for P. falciparum. It allowed determining parasitemia levels of 0.1% or higher, with coefficient of variation smaller than 10%. In terms of the precision range of parasitemia, both high and low ranges showed similar precision results. Pearson's correlation test was used to evaluate the correlation data coming from all methods. A strong correlation of measured parasitemia (r2=0.99, P<0.05) was observed between each method. The parasitemia analysis using this new diagnostic tool needs technical improvement, particularly in the differentiation of malaria species.

Isolation and identification of mammalian orthoreovirus type 3 from a Korean roe deer (Capreolus pygargus)

  • Yang, Dong-Kun;An, Sungjun;Park, Yeseul;Yoo, Jae Young;Park, Yu-Ri;Park, Jungwon;Kim, Jong-Taek;Ahn, Sangjin;Hyun, Bang-Hun
    • 대한수의학회지
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    • 제61권2호
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    • pp.13.1-13.8
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    • 2021
  • Mammalian reovirus (MRV) causes respiratory and intestinal disease in mammals. Although MRV isolates have been reported to circulate in several animals, there are no reports on Korean MRV isolates from wildlife. We investigated the biological and molecular characteristics of Korean MRV isolates based on the nucleotide sequence of the segment 1 gene. In total, 144 swabs from wild animals were prepared for virus isolation. Based on virus isolation with specific cytopathic effects, indirect fluorescence assays, electron microscopy, and reverse transcription-polymerase chain reaction, only one isolate was confirmed to be MRV from a Korean roe deer (Capreolus pygargus). The isolate exhibited a hemagglutination activity level of 16 units with pig erythrocytes and had a maximum viral titer of 105.7 50% tissue culture infectious dose (TCID50)/mL in Vero cells at 5 days after inoculation. The nucleotide and amino-acid sequences of the partial segment S1 of the MReo2045 isolate were determined and compared with those of other MRV strains. The MReo2045 isolate had nucleotide sequences similar to MRV-3 and was most similar (96.1%) to the T3/Bat/Germany/342/08 strain, which was isolated in Germany in 2008. The MReo2045 isolate will be useful as an antigen for sero-epidemiological studies and developing diagnostic tools.

제주 스코리아로부터 합성한 제올라이트 물질에 의한 암모니아성 질소의 흡착 특성 (Adsorption Characteristics of Ammonia-Nitrogen by Zeolitic Materials Synthesized from Jeju Scoria)

  • 이창한;현성수;감상규
    • 한국환경과학회지
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    • 제29권12호
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    • pp.1261-1274
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    • 2020
  • The characteristics of ammonia-nitrogen (NH4+-N) adsorption by a zeolitic material synthesized from Jeju scoria using the fusion and hydrothermal method was studied. The synthetic zeolitic material (Z-SA) was identified as a Na-A zeolite by X-ray diffraction, X-ray fluorescence analysis and scanning electron microscopy images. The adsorption of NH4+-N using Jeju scoria and different types of zeolite such as the Z-SA, natural zeolite, and commercial pure zeolite (Na-A zeolite, Z-CS) was compared. The equilibrium of NH4+-N adsorption was reached within 30 min for Z-SA and Z-CS, and after 60 min for Jeju scoria and natural zeolite. The adsorption capacity of NH4+-N increased with approaching to neutral when pH was in the range of 3-7, but decreased above 7. The removal efficiency of NH4+-N increased with increasing Z-SA dosage, however, its adsorption capacity decreased. For initial NH4+-N concentrations of 10-200 mg/L at pH 7, the adsorption rate of NH4+-N was well described by the pseudo second-order kinetic model than the pseudo first-order kinetic model. The adsorption isotherm was well fitted by the Langmuir model. The maximum uptake of NH4+-N obtained from the Langmuir model decreased in the order of Z-CS (46.8 mg/g) > Z-SA (31.3 mg/g) > natural zeolite (5.6 mg/g) > Jeju scoria (0.2 mg/g).